• Korean Journal of Veterinary Research
• /
• v.34 no.4
• /
• pp.781-785
• /
• 1994

Uridine diphosphate(UDP)-galactose-4-epimerase-less mutants of Salmonella pullorum were isolated after mutagenic treatment with ethidium bromide. When isolated gal E mutants of S. pullorum A2 and D1 were grown in the presence of galactose(0.1 W/V), they exhibited marked bacteriolysis in heart infusion broth. The mutant strains were further investigated the characteristics of enzymatic activities in the Leoloir galactose pathway. Isolated A2 and D1 strains were completely deficient in UDP-galactose-4-epimerase activity. And the activity of other enzymes involved in galactose metabolism were reduced significantly.

• Journal of Microbiology and Biotechnology
• /
• v.18 no.8
• /
• pp.1439-1444
• /
• 2008

The recombinant E. coli $\Delta$6 mutant (galR, glpK, gldA, IdhA, lacI, tpiA) was used to produce 1,3-propanediol (PD) from glucose. The 1,3-PD production increased with feedback control of the glucose concentration using fed-batch fermentation. The maximum 1,3-PD concentration produced was 43 g/l after 60 h of fermentation. Glycerol production was minimized when controlling the glucose concentration at less than 1 g/l. The expression levels of seven enzymes related to the 1,3-PD production metabolism were compared during the cell growth phase and 1,3-PD production phase, and their expression levels all increased during 1,3-PD production, with the exception of alcohol dehydrogenase.

• Reproductive and Developmental Biology
• /
• v.34 no.1
• /
• pp.33-40
• /
• 2010

Equine chorionic gonadotropin (eCG) is a heavily glycosylated glycoprotein composed of non-covalently linked $\alpha$- and $\beta$-subunits. To study the function and signal transduction of tethered recombinant-eCG (rec-eCG), a single chain eCG molecule was constructed, and the rec-eCG protein was prepared. In this study, we constructed 5 mutants (${\Delta}1$, ${\Delta}2$, ${\Delta}3$, ${\Delta}4$, and ${\Delta}5$) of rec-eCG using data about known glycoprotein hormones to analyze the role of specific follicle stimulating homone (FSH)-like activity. Three amino acids of certain specific sites were replaced with alanine. The expression vectors were transfected into CHO cells and subjected to G418 selection for 2~3 weeks. The media were collected and the quantity of secreted tethered rec-eCGs was quantified by ELISA. The LH- and FSH-like activities were assayed in terms of cAMP production by rat LH/CG and rat FSH receptors. Then, the metabolic clearance rate analyzed by the injection of rec-eCG (5 IU) into the tail vein was analyzed. The mutant eCGs (${\Delta}l$, ${\Delta}4$, and ${\Delta}5$) were transcripted, but not translated into proteins. Rec-eCG A2 was secreted in much lower amounts than the wild type. Only the rec-eCG ${\Delta}3$ ($\beta$-subunit: $Gln^{94}-Ile^{95}-Lys^{96}{\rightarrow}Ala^{94}-Ala^{95}-Ala^{96}$) was efficiently secreted. Although activity is low, its LH-like activity was similar to that of tethered $eCG{\beta\alpha}$. However, the FSH-like activity of rec-$eCG{\beta\alpha\Delta}3$ was completely flat. The result of the analysis of the metabolic clearance rate shoed the persistence of the mutant in the blood until 4 hours after the injection. After then, it almost disappeared at 8 hours. Taken together, these data suggest that 94~96 amino acid sequences in eCG $\beta$-subunit appear to be of utmost importance for signal transduction of the FSH receptor.